Poly-β-hydroxybutyrate (PHB) is a biodegradable polymer, synthesized as carbon and energy reserve by bacteria and archaea. To the best of our knowledge, this is the first report on PHB production by a rare actinomycete species, Rhodococcus pyridinivorans BSRT1-1. Response surface methodology (RSM) employing central composite design, was applied to enhance PHB production in a flask scale. A maximum yield of 3.6 ± 0.5 g/L in biomass and 43.1 ± 0.5 wt% of dry cell weight (DCW) of PHB were obtained when using RSM optimized medium, which was improved the production of biomass and PHB content by 2.5 and 2.3-fold, respectively. The optimized medium was applied to upscale PHB production in a 10 L stirred-tank bioreactor, maximum biomass of 5.2 ± 0.5 g/L, and PHB content of 46.8 ± 2 wt% DCW were achieved. Furthermore, the FTIR and 1H NMR results confirmed the polymer as PHB. DSC and TGA analysis results revealed the melting, glass transition, and thermal decomposition temperature of 171.8, 4.03, and 288 °C, respectively. In conclusion, RSM can be a promising technique to improve PHB production by a newly isolated strain of R. pyridinivorans BSRT1-1 and the properties of produced PHB possessed similar properties compared to commercial PHB.

This study focused on decolorization of 2 reactive dyes; Reactive Blue 19 (RBBR) and Reactive Black 5 (RB5), by selected white-rot fungus Datronia sp. KAPI0039. The effects of reactive dye concentration, fungal inoculum size as well as pH were studied. Samples were periodically collected for the measurement of color unit, Laccase (Lac), Manganese Peroxidase (MnP), and Lignin Peroxidase (LiP) activity. Eighty-six percent of 1,000 mg L −1 RBBR decolorization was achieved by 2% (w/v) Datronia sp. KAPI0039 at pH 5. The highest Lac activity (759.81 UL −1 ) was detected in the optimal condition. For RB5, Datronia sp. KAPI0039 efficiently performed (88.01% decolorization) at 2% (w/v) fungal inoculum size for the reduction of 600 mg L −1 RB5 under pH 5. The highest Lac activity (178.57 UL −1 ) was detected, whereas the activity of MnP and LiP was absent during this hour. The result, therefore, indicated that Datronia sp. KAPI0039 was obviously able to breakdown both reactive dyes, and Lac was considered as a major lignin-degradation enzyme in this reaction.

In order to develop a method for producing fuel ethanol from cassava pulp using cell surface engineering (arming) technology, an arming yeast co-displaying α-amylase (α-AM), glucoamylase, endoglucanase, cellobiohydrase, and β-glucosidase on the surface of the yeast cells was constructed. The novel yeast strain, possessing the activities of all enzymes, was able to produce ethanol directly from soluble starch, barley β-glucan, and acid-treated Avicel. Cassava is a major crop in Southeast Asia and used mainly for starch production. In the starch manufacturing process, large amounts of solid wastes, called cassava pulp, are produced. The major components of cassava pulp are starch (approximately 60%) and cellulose fiber (approximately 30%). We attempted simultaneous saccharification and ethanol fermentation of cassava pulp with this arming yeast. During fermentation, ethanol concentration increased as the starch and cellulose fiber substrates contained in the cassava pulp decreased. The results clearly showed that the arming yeast was able to produce ethanol directly from cassava pulp without addition of any hydrolytic enzymes.

Bromelain is a mixture of proteolytic enzymes derived from pineapple (Ananas comosus) fruit and stem possessing several beneficial properties, particularly anti-inflammatory activity. However, the molecular mechanisms underlying the anti-inflammatory effects of bromelain are unclear. This study investigated the anti-inflammatory effects and inhibitory molecular mechanisms of crude and purified rhizome bromelains on lipopolysaccharide (LPS)-induced inflammation in RAW 264.7 macrophage cells. RAW264.7 cells were pre-treated with various concentrations of crude bromelain (CB) or purified bromelain (PB), and then treated with LPS. The production levels of pro-inflammatory cytokines and mediators, including nitric oxide (NO), interleukin (IL)-6, and tumor necrosis factor (TNF)-α were determined by Griess and ELISA assays. The expressions of inducible nitric oxide synthetase (iNOS), cyclooxygenase (COX)-2, nuclear factor kappa B (NF-κB), and mitogen-activated protein kinases (MAPKs)-signaling pathway-related proteins were examined by western blot analysis. The pre-treatment of bromelain dose-dependently reduced LPS-induced pro-inflammatory cytokines and mediators, which correlated with downregulation of iNOS and COX-2 expressions. The inhibitory potency of PB was stronger than that of CB. PB also suppressed phosphorylated NF-κB (p65), nuclear factor of kappa light polypeptide gene enhancer in B-cells inhibitor alpha, extracellular signal-regulated kinases, c-Jun amino-terminal kinases, and p38 proteins in LPS-treated cells. PB then exhibited potent anti-inflammatory effects on LPS-induced inflammatory responses in RAW264.7 cells by inhibiting the NF-κB and MAPKs-signaling pathways.

β-Glucan (BG), one of the most abundant polysaccharides containing glucose monomers linked by β-glycosidic linkages, is prevalent in yeast biomass that needs to be recovered to obtain this valuable polymer. This study aimed to apply alkaline and enzymatic processes for the recovery of BG from the yeast strain Kluyveromyces marxianus TISTR 5925. For this purpose, the yeast was cultivated to produce the maximum yield of raw material (yeast cells). The effective recovery of BG was then established using either an alkaline or an enzymatic process. BG recovery of 35.45% was obtained by using 1 M NaOH at 90 °C for 1 h, and of 81.15% from 1% (w/v) hydrolytic protease enzyme at 55 °C for 5 h. However, BG recovered by the alkaline process was purer than that obtained by the enzymatic process. Fourier transform infrared (FTIR) and nuclear magnetic resonance (NMR) spectroscopy confirmed the purity, the functional groups, and the linkages of BG obtained from different recovery systems and different raw materials. The results of this study suggest that an alkaline process could be an effective approach for the solubilization and recovery of considerable purity of BG from the yeast cells. In addition, the obtained BG had comparable functional properties with commercially available BG. This study reveals the effectiveness of both chemical and biological recovery of BG obtained from yeast as a potential polymeric material.

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Polyhydroxyalkanoates (PHAs) are biodegradable polymers synthesized by certain bacteria and archaea with functions comparable to conventional plastics. Previously, our research group reported a newly PHA-producing bacterial strain, Rhodococcus pyridinivorans BSRT1-1, from the soil in Thailand. However, this strain’s PHA synthase (phaCRp) gene has not yet been characterized. Thus, this study aims to synthesize PHA using a newly engineered bacterial strain, Cupriavidus necator PHB−4/pBBR_CnPro-phaCRp, which harbors the phaCRp from strain BSRT1-1, and characterize the properties of PHA for skin tissue engineering application. To the best of our knowledge, this is the first study on the characterization of the PhaC from R. pyridinivorans species. The results demonstrated that the expression of the phaCRp in C. necator PHB−4 had developed in PHA production up to 3.1 ± 0.3 g/L when using 10 g/L of crude palm kernel oil (CPKO) as a sole carbon source. Interestingly, the engineered strain produced a 3-hydroxybutyrate (3HB) with 2 mol% of 3-hydroxyhexanoate (3HHx) monomer without adding precursor substrates. In addition, the 70 L stirrer bioreactor improved P(3HB-co-2 mol% 3HHx) yield 1.4-fold over the flask scale without altering monomer composition. Furthermore, the characterization of copolymer properties showed that this copolymer is promising for skin tissue engineering applications.

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